甲酸夹层法非常适合丝状真菌鉴定，并通过 Zybio EXS2600 质谱提高了周转时间。
Formic acid sandwich method is well-suited for filamentous fungi identification and improves turn around time using Zybio EXS2600 mass spectrometry.
机构信息
Department of Laboratory Medicine, West China Hospital of Sichuan University, Sichuan Province, Chengdu, 610041, P.R. China.
Department of Laboratory Medicine, Yaan People's Hospital, Yaan, 625000, China.
出版信息
BMC Microbiol. 2024 Jul 3;24(1):238. doi: 10.1186/s12866-024-03394-2.
OBJECTIVES
Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is extensively employed for the identification of filamentous fungi on MALDI Biotyper (Bruker Daltonics) and Vitek MS (biomerieux), but the performance of fungi identification on new EXS2600 (Zybio) is still unknow. Our study aims to evaluate the new EXS2600 system's (Zybio) ability to rapidly identify filamentous fungi and determine its effect on turnaround time (TAT) in our laboratory.
METHODS
We tested 117 filamentous fungi using two pretreatment methods: the formic acid sandwich (FA-sandwich) and a commercial mold extraction kit (MEK, Zybio). All isolates were confirmed via sequence analysis. Laboratory data were extracted from our laboratory information system over two 9-month periods: pre-EXS (April to December 2022) and post-EXS (April to December 2023), respectively.
RESULTS
The total correct identification (at the species, genus, or complex/group level) rate of fungi was high, FA-sandwich (95.73%, 112/117), followed by MEK (94.02%, 110/117). Excluding 6 isolates not in the database, species-level identification accuracy was 92.79% (103/111) for FA-sandwich and 91.89% (102/111) for MEK; genus-level accuracy was 97.29% (108/111) and 96.39% (107/111), respectively. Both methods attained a 100% correct identification rate for Aspergillus, Lichtheimia, Rhizopus Mucor and Talaromyces species, and were able to differentiate between Fusarium verticillioides and Fusarium proliferatum within the Fusarium fujikuroi species complex. Notably, high confidence was observed in the species-level identification of uncommon fungi such as Trichothecium roseum and Geotrichum candidum. The TAT for all positive cultures decreased from pre EXS2600 to post (108.379 VS 102.438, P < 0.05), and the TAT for tissue decreased most (451.538 VS 222.304, P < 0.001).
CONCLUSIONS
The FA-sandwich method is more efficient and accurate for identifying filamentous fungi with EXS2600 than the MEK. Our study firstly evaluated the performance of fungi identification on EXS2600 and showed it is suitable for clinical microbiology laboratories use.
目的
基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）广泛用于 MALDI Biotyper（布鲁克·道尔顿）和 Vitek MS（生物梅里埃）上丝状真菌的鉴定，但新型 EXS2600（Zybio）上真菌鉴定的性能仍不清楚。本研究旨在评估新型 EXS2600 系统（Zybio）快速鉴定丝状真菌的能力，并确定其对我们实验室周转时间（TAT）的影响。
方法
我们使用两种预处理方法测试了 117 株丝状真菌：甲酸夹层（FA-sandwich）和商用霉菌提取试剂盒（MEK，Zybio）。所有分离株均通过序列分析确认。实验室数据从我们的实验室信息系统中提取，时间跨度为两个 9 个月：前 EXS（2022 年 4 月至 12 月）和后 EXS（2023 年 4 月至 12 月）。
结果
真菌的总正确鉴定率（种、属或复合体/组水平）很高，FA-sandwich（95.73%，112/117），其次是 MEK（94.02%，110/117）。排除 6 株不在数据库中的菌株，FA-sandwich 的种水平鉴定准确率为 92.79%（103/111），MEK 为 91.89%（102/111）；属水平的准确率分别为 97.29%（108/111）和 96.39%（107/111）。两种方法均能 100%正确鉴定曲霉属、亮菌属、根霉属、毛霉属和拟青霉属的种，且能够区分镰孢菌属 verticillioides 和 fusarium proliferatum 种复合体中的 fusarium fujikuroi 种。值得注意的是，在鉴定 Trichothecium roseum 和 Geotrichum candidum 等不常见真菌时，鉴定的置信度很高。所有阳性培养物的 TAT 从预 EXS2600 到后均降低（108.379 对 102.438，P<0.05），组织的 TAT 降低最多（451.538 对 222.304，P<0.001）。
结论
FA-sandwich 法比 MEK 法更高效、准确地鉴定丝状真菌。本研究首次评估了 EXS2600 上真菌鉴定的性能，表明其适用于临床微生物学实验室使用。
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